Injectable ready-to-use solutions containing an antitumor anthracycline glycoside

ABSTRACT

According to the invention there is provided a sterile, pyrogen-free, ready-to-use solution of an anthracycline glycoside, especially doxorubicin, which consists essentially of a physiologically acceptable salt of an anthracycline glycoside dissolved in a physiologically acceptable solvent therefor, which has not been reconstituted from a lyophilizate and which has a pH of from 2.5 to 6.5. The solution of the invention is particularly advantageous for the administration by injection of the anthracycline glycoside drugs, e.g. doxorubicin, in the treatment of both human and animal tumors.

This is a continuation application of U.S. application Ser. No.09/149,360, filed Sep. 8, 1998 now U.S. Pat. No. 6,284,738, which is acontinuation application of U.S. application Ser. No. 07/827,742, filedJan. 29, 1992 now U.S. Pat. No. 6,107,285, which is a divisionalapplication of U.S. Ser. No. 07/503,856, filed Apr. 3, 1990 now U.S.Pat. No. 5,124,317, which is a divisional application of U.S. Ser. No.07/385,999, filed Jul. 27, 1989 now U.S. Pat. No. 4,946,831, which inturn is a continuation application of U.S. Ser. No. 06/878,784, filedJun. 26, 1986, now abandoned.

The present invention relates to a stable intravenously injectableready-to-use solution of an antitumor anthracycline glycoside, e.g.doxorubicin, to a process for preparing such a solution, and provide thesame in a sealed container, and to a method for treating tumors by theuse of the said ready-to-use solution.

The anthracycline glycoside compounds are a well known class ofcompounds in the antineoplastic group of agents, wherein doxorubicin isa typical, and the most widely used, representative: Doxorubicin.Anticancer Antibiotics, Federico Arcamone, 1981, Publ: Academic Press,New York, N.Y.; Adriamycin Review, EROTC International Symposium,Brussels, May, 1974, edited by M. Staquet. Publ. Eur. Press Medikon,Ghent, Belg.; Results of Adriamycin Therapy, Adriamycin Symposium atFrankfurt/Main 1974 edited by M. Ghione, J. Fetzer and H. Maier, publ.:Springer, New York, N.Y.

At present, anthracycline glycoside antitumor drugs, in particular,e.g., doxorubicin, are solely available in the form of lyophilizedpreparations, which need to be reconstituted before administration. Boththe manufacturing and the reconstitution of such preparations expose theinvolved personnel (workers, pharmacists, medical personnel, nurses) torisks of contamination which are particularly serious due to thetoxicity of the antitumor substances. The Martindale Extra Pharmacopoeia28th edition, page 175 left column, reports, indeed, about adverseeffects of antineoplastic drugs and recommends that “They must behandled with great care and contact with skin and eyes avoided; theyshould not be inhaled. Care must be taken to avoid extravasation sincepain and tissue damage may ensue.”

Similarly, Scand. J. Work Environ Health vol. 10 (2), pages 71-74(1984), as well as articles on Chemistry Industry, Issue Jul. 4, 1983,page 488, and Drug-Topics-Medical-Economics-Co, Issue Feb. 7, 1983, page99 report about severe adverse effects observed in medical personnelexposed to use of cytostatic agents, including doxorubicin.

To administer a lyophilized preparation, double handling of the drug isrequired, the lyophilized cake having to be first reconstituted and thenadministered and, moreover, in some cases, the complete dissolution ofthe powder may require prolonged shaking because of solubilizationproblems.

As the risks connected with the manufacturing and the reconstitution ofa lyophilized preparate would be highly reduced if a ready-to-usesolution of the drug were available, we have developed a stable,therapeutically acceptable intravenously injectable solution of ananthracycline glycoside drug, e.g. doxorubicin, whose preparation andadministration does not require either lyophilization or reconstitution.

According to the present invention, there is provided a sterile,pyrogen-free, anthracycline glycoside solution which consistsessentially of a physiologically acceptable salt of an anthracyclineglycoside dissolved in a physiologically acceptable solvent therefor,which has not been reconstituted from a lyophilizate and which has a pHof from 2.5 to 6.5.

Preferably the solution of the invention is provided in a sealedcontainer.

Preferably the anthracycline glycoside is chosen from the groupconsisting of doxorubicin, 4′-epi-doxorubicin (i.e. epirubicin),4′-desoxy-doxorubicin (i.e. esorubicin), 4′-desoxy-4′-iodo-doxorubicin,daunorubicin and 4-demethoxydaunorubicin (i.e. idarubicin).

A particularly preferred anthracycline glycoside is doxorubicin.

Any physiologically acceptable salt of the anthracycline glycoside maybe used for preparing the solution of the invention. Examples ofsuitable salts may be, for instance, the salts with mineral inorganicacids such as hydrochloric, hydrobromic, sulfuric, phosphoric, nitricand the like, and the salts with certain organic acids such as acetic,succinic, tartaric, ascorbic, citric, glutammic, benzoic,methanesulfonic, ethanesulfonic and the like. The salt with hydrochloricacid is a particularly preferred salt, especially when the anthracyclineglycoside is doxorubicin.

Any solvent which is physiologically acceptable and which is able todissolve the anthracycline glycoside salt may be used. The solution ofthe invention may also contain one or more additional components such asa co-solubilizing agent (which may be the same as a solvent), a tonicityadjustment agent and a preservative. Examples of solvents,co-solubilizing agents, tonicity adjustment agents and preservativeswhich can be used for the preparation of the anthracycline glycosidesolutions of the invention are hereunder reported.

Suitable solvents and co-solubilizing agents may be, for instance,water; physiological saline; aliphatic amides, e.g.N,N-dimethylacetamide, N-hydroxy-2-ethyl-lactamide and the like;alcohols, e.g. ethanol, benzyl alcohol and the like; glycols andpolyalcohols, e.g. propyleneglycol, glycerin and the like; esters ofpolyalcohols, e.g. diacetine, triacetine and the like; polyglycols andpolyethers, e.g. polyethyleneglycol 400, propyleneglycol methylethersand the like; dioxolanes, e.g. isopropylidenglycerin and the like;dimethylisosorbide; pyrrolidone derivatives, e.g. 2-pyrrolidone,N-methyl-2-pyrrolidone, polyvinylpyrrolidone (co-solubilizing agentonly) and the like; polyoxyethylenated fatty alcohols, e.g. Brij® andthe like; esters of polyoxyethylenated fatty acids, e.g. Cremophor®,Myij® and the like; polysorbates, e.g. Tweens®; polyoxyethylenederivatives of polypropyleneglycols, e.g. Pluronics®.

A particularly preferred co-solubilizing agent is polyvinylpyrrolidone.

Suitable tonicity adjustment agents may be, for instance,physiologically acceptable inorganic chlorides, e.g. sodium chloride,dextrose, lactose, mannitol and the like.

Preservatives suitable for physiological administration may be, forinstance, esters of para-hydroxybenzoic acid (e.g., methyl, ethyl,propyl and butyl esters, or mixtures of them), chlorocresol and thelike.

The above mentioned solvents and co-solubilizing agents, tonicityadjustment agents and preservatives can be used alone or as a mixture oftwo or more of them.

Examples of preferred solvents are water, ethanol, polyethyleneglycoland dimethylacetamide as well as mixtures in various proportions ofthese solvents. Water is a particularly preferred solvent.

To adjust the pH within the range of from 2.5 to about 5.0 aphysiologically acceptable acid may be added as desired. The acid may beany physiologically acceptable acid, e.g., an inorganic mineral acidsuch as hydrochloric, hydrobromic, sulfuric, phosphoric, nitric and thelike, or an organic acid such as acetic, succinic, tartaric, ascorbic,citric, glutammic, benzoic, methanesulphonic, ethanesulfonic and thelike, or also an acidic physiologically acceptable buffer solution,e.g., a chloride buffer, an acetate buffer, a phosphate buffer and thelike.

For obtaining pH values from about 5 to about 5.5 the addition of theacid is not, usually, necessary, but only addition of a physiologicallyacceptable buffer solution, e.g., one of those indicated above, may berequired, as desired.

For obtaining pH values from about 5.5 to 6.5 the addition of aphysiologically acceptable alkalinizing agent, such as sodium hydroxide,a mono, di- or triethanolamine or the like, or, preferably, a buffersolution such as a phosphate buffer, a TRIS buffer or the like isrequired.

The preferred range of pH for the ready-to-use solution of the inventionis from 2.5 to 5.5, in particular from about 3 to about 5.2, a pH ofabout 3 and a pH of about 5 being particularly preferred values.

In the solutions of the invention the concentration of the anthracyclineglycoside may vary within broad ranges, preferably from 0.1 mg/ml to 100mg/ml, in particular from 0.1 mg/ml to 50 mg/ml, most preferably from 1mg/ml to 20 mg/ml.

The preferred ranges of concentration may be slightly different fordifferent anthracycline glycosides. Thus, for example, preferredconcentrations for doxorubicin are from about 2 mg/ml to about 50 mg/ml,preferably from 2 mg/ml to 20 mg/ml, particularly appropriate valuesbeing 2 mg/ml and 5 mg/ml. Similar concentrations are preferred also for4′-epi-doxorubicin, 4′-desoxy-doxorubicin and4′-desoxy-4′-iodo-doxorubicin. Preferred ranges of concentration fordaunorubicin and 4-demethoxy-daunorubicin are from 0.1 mg/ml to 50mg/ml, preferably from 1 mg/ml to 20 mg/ml, concentrations of 1 mg/mland 5 mg/ml being particularly appropriate.

Suitable packaging for the anthracycline glycoside solutions may be allapproved containers intended for parenteral use, such as plastic andglass containers, ready-to-use syringes and the like. Preferably thecontainer is a sealed glass container, e.g. a vial or an ampoule.

According to a particularly preferred feature of the invention, there isprovided a sterile, pyrogen-free, doxorubicin solution which consistsessentially of a physiologically acceptable salt of doxorubicindissolved in a physiologically acceptable solvent therefor, which hasnot been reconstituted from a lyophilizate and which has a pH of from2.5 to 6.5.

In the above indicated preferred feature of the invention thephysiologically acceptable salt of doxorubicin may be, e.g. the saltwith a mineral inorganic acid such as hydrochloric, hydrobromic,sulfuric, phosphoric, nitric and the like, or the salt with an organicacid such as acetic, succinic, tartaric, ascorbic, citric, glutammic,benzoic, methanesulfonic, ethanesulfonic and the like. The hydrochloridesalt is a particularly preferred salt.

For the solution hereabove indicated as a preferred feature of theinvention suitable solvents, co-solubilizing agents, tonicity adjustmentagents and preservatives may be the same as those previously recited inthe specification. Water is a particularly preferred solvent.

Also, the physiologically acceptable acid which may be added to adjustthe pH to from 2.5 to about 5, if desired, and the alkanilizing agentwhich may be added to adjust the pH, if desired, to a value from about5.5 to 6.5 may be one of those previously specified. When it is desiredto adjust the pH of the above said preferred solution to a value of from2.5 to about 5, hydrochloric acid is an especially preferred acid.Preferred pH values for the above said preferred solutions of theinvention are from 2.5 to 5.5, in particular from about 3 to about 5.2,the pH values of 3 and 5 being especially preferred.

Though the concentration of doxorubicin in the above preferred featuremay vary within the broad range from 0.1 mg/ml to 100 mg/ml, preferredconcentrations are from 2 mg/ml to 50 mg/ml, most preferably from 2mg/ml to 20 mg/ml: examples of especially preferred concentrations ofdoxorubicin are 2 mg/ml and 5 mg/ml.

The invention also provides a process for producing a sterile,pyrogen-free anthracycline glycoside solution with a pH of from 2.5 to6.5, which process comprises dissolving a physiologically acceptablesalt of the anthracycline glycoside, which salt is not in the form of alyophilizate, in a physiologically acceptable solvent therefor;optionally adding a physiologically acceptable acid or buffer to adjustthe pH within the said range as desired; and passing the resultingsolution through a sterilising filter.

One or more additional components such as co-solubilizing agents,tonicity adjustment agents and preservatives, for instance of the kindpreviously specified, may be added to the solution prior to passing thesolution through the sterilising filter.

With the solutions of the invention it is possible to obtaincompositions having a very high concentration of the anthracyclineglycoside active substance even at 50 mg/ml and more. This constitutes agreat advantage over the presently available lyophilized preparateswherein high concentrations of anthracycline glycoside can only beobtained with difficulty because of solubilization problems encounteredin reconstitution, mainly with saline. The presence of the excipient,e.g. lactose, in the lyophilized cake, and its generally high proportionin respect of the active substance, even up to 5 parts of excipient perpart of active substance, has a negative effect on solubilization sothat difficulties may arise in obtaining dissolution of the lyophilizedcake, especially for concentrations of anthracycline glycoside higherthan 2 mg/ml.

The solutions of the invention are characterized by a good stability.Solutions in various solvents and with different pH's and concentrationshave been found to be stable for long periods at temperatures acceptedfor the storage of pharmaceutical preparations. This is illustrated inthe Examples which follow.

Owing to the well known anti-tumor activity of the anthracyclineglycoside active drug substance, the pharmaceutical compositions of theinvention are useful for treating tumors in both human and animal hosts.Examples of tumors that can be treated are, for instance, sarcomas,including osteogenic and soft tissue sarcomas, carcinomas, e.g.,breast-, lung-, bladder-, thyroid-, prostate- and ovarian carcinoma,lymphomas, including Hodgkin and non-Hodgkin lymphomas, neuroblastoma,melanoma, myeloma, Wilms tumor, and leukemias, including acutelymphoblastic leukemia and acute myeloblastic leukemia. Examples ofspecific tumours that can be treated are Moloney Sarcoma Virus, Sarcoma180 Ascites, solid Sarcoma 180, gross transplantable leukemia, L 1210leukemia and lymphocytic P 368 leukemia.

Thus, according to the invention there is also provided a method ofinhibiting the growth of a tumour, in particular one of those indicatedabove, which comprises administering to a host suffering from saidtumour an injectable solution according to the invention containing theactive drug substance in an amount sufficient to inhibit the growth ofsaid tumour.

The injectable solutions of the invention are administered by rapidintravenous injection or infusion according to a variety of possibledose schedules. Suitable dose schedule for doxorubicin may be, forexample, of 60 to 75 mg of active drug substance per m² of body surfacegiven as a single rapid infusion and repeated at 21 days; an alternativeschedule may be of 30 mg/m² day by intravenous route for 3 days, every25 days. Suitable dosages for 4′-epi-doxorubicin and4′-desoxy-doxorubicin may be, for instance, of 75 to 90 mg/m² given in asingle infusion to be repeated at 21 days, and similar dosages may beuseful for 4′-desoxy-4′-iodo-doxorubicin.

Idarubicin, i.e. 4-demethoxy-daunorubicin, may be, e.g., administeredintravenously at a single dose of 13-15 mg/m² every 21 days in thetreatment of solid tumours, while in the treatment of leukemias apreferred dose schedule is, e.g., of 10-12 mg/m² day by intravenousroute for 3 days, to be repeated every 15-21 days; similar dosages maybe, e.g., followed also for daunorubicin.

The following examples illustrate but do not limit in any way theinvention.

With reference to the examples, the stability controls on theready-to-use solutions were carried out by means of high performanceliquid chromatography (HPLC), at the following experimental conditions:

Liquid chromatograph : Varian model 5010 Spectrophotometric detector :Knauer model 8700 Integrating recorder : Varian model CDS 401 Injectionvalve : Rheodyne model 7125 fitted with a 10 mcl sample loopChromatographic column : Waters μ-Bondapak C18 (length = 300 mm; innerdiameter = 3.9 mm; average particle size = 10 mcm) Column temperature :ambient (about 22° C. ± 2° C.) Mobile phase : water: acetonitrile (69:31v/v) adjusted top pH 2 with phosphoric acid, filtered (sintered glassfilter, 1 mcm or finer porosity) and deaerated Mobile phase flow rate :1.5 ml/min Analytical wavelength : 254 ± 1 nm Integrating recordersensitivity : 512 Chart speed : 1 cm/min

At these conditions, the peak of the anthracycline glycoside showed aretention time of about 6 minutes.

The obtained results are reported in the Tables accompanying theexamples.

The extrapolation of the analytical data in order to determine the timewhen the 90% of the initial assay could be expected (t₉₀ value) was madefollowing an Arrhenius plot.

This procedure of analytical data treatment is well known and widelyused and described in the art: see, e.g., Chemical Stability ofPharmaceuticals, Kennet A. Connors, Gordon L. Amidon, Lloyd Kennon,Publ. John Wiley and Sons, New York, N.Y., 1979.

The term “teflon” refers to “Teflon™”.

EXAMPLE 1

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.8 g (10 mg)Water for injections 0.4 l  (5 ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The pH of thesolution was not adjusted. Further de-aerated water for injections wasthen added to bring the solution to its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 1:

TABLE 1 INITIAL VALUES Concentration: 1.994 mg/ml   pH = 5.2 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.992 99.9 1.917 96.1 1.768 88.7 1.493 75.0 21.843 92.4 1.618 81.1 1.166 58.5 3 1.774 89.0 1.506 75.5 0.830 41.6 41.974 99.0 1.720 86.3 1.393 69.9 12 1.980 99.3 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 815 days t₉₀ at 8° C. =480 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 2

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.8 g (10 mg)Hydrochloric acid 0.1N pH = 3 (pH = 3) q.s. to Water for injections 0.4l  (5 ml) q.s. to

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 2:

TABLE 2 INITIAL VALUES Concentration: 1.992 mg/ml   pH = 3.0 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.995 100.2 1.952 98.0 1.919 96.3 1.493 75.0 21.889 94.8 1.851 92.9 1.036 51.9 3 1.876 94.2 1.565 78.6 0.730 36.7 41.979 99.4 1.808 90.8 1.393 69.9 12 1.972 99.0 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 3970 days t₉₀ at 8° C. =2000 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 3

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 8.0 g (100 mg)Hydrochloric acid 0.1N pH = 3 (pH = 3) q.s. to Water for injections 0.4l  (5 ml) q.s. to

Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials ere thenclosed with chlorobutyl teflon-faced rubber stoppers and sealed withaluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatogrpahy (HPLC) for the determination of potency, are reported inthe following Table 3:

TABLE 3 INITIAL VALUES Concentration: 20.06 mg/ml   pH = 2.95 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 20.06 100.0 19.56 97.5 17.84 88.9 12.31 61.4 218.87 94.1 15.61 77.8 7.09 35.3 3 18.24 90.9 13.41 66.8 3.13 15.6 419.91 99.2 17.51 87.3 11.07 55.2 12 19.80 98.7 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 3700 days t₉₀ at 8° C. =1780 days

Similar stability data can be observed also for analogous solutionscontaining 4′-epi-doxorubicin or 4′-desoxydoxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 4

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.80 (10.0 mg)Polyvinylpyrrolidone 20.00 g (250.0 mg) Water for injections 0.40 l (5.0ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the water forinjections, de-aerated by nitrogen bubbling. The pH of the solution wasnot adjusted. Polyvinylpyrrolidone was added and dissolved understirring and nitrogen bubbling. Further de-aerated water for injectionswas then added to bring the solution to its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 4:

TABLE 4 INITIAL VALUES Concentration: 1.986 mg/ml   pH = 4.6 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.984 99.9 1.928 97.1 1.797 90.5 1.605 80.8 21.847 93.0 1.616 81.4 1.293 65.1 3 1.828 92.0 1.527 76.9 1.018 51.3 41.928 97.1 1.797 90.5 1.403 70.7 8 1.989 100.1 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 1460 days t₉₀ at 8° C. =835 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 5

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.800 g (10.00mg) N,N-Dimethylacetamide 0.060 l (0.75 ml) Propylene glycol 0.048 l(0.60 ml) Ethanol 0.012 l (0.15 ml) Hydrochloric acid 0.1N pH = 3 (pH =3) q.s. to Water for injections 0.400 l (5.00 ml) q.s. to

Doxorubicin.HCl (0.800 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-Dimethylacetamide, propylene glycol and ethanol were subsequentlyadded under stirring and nitrogen bubbling. The hydrochloric acid wasthen added dropwise to adjust the pH of the solution to 3. Furtherde-aerated water for injections was then added to bring the solution toits final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 5:

TABLE 5 INITIAL VALUES Concentration: 2.000 mg/ml   pH = 3.03 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.892 94.6 1.735 86.7 1.495 74.7 2 1.993 99.71.927 96.4 1.624 81.2 1.212 60.6 3 1.908 95.4 1.432 71.6 1.032 51.6 42.00 100.0 1.863 93.2 1.266 63.3 8 1.960 98.0 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 4360 days t₉₀ at 8° C. =2200 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 6

for Composition 80 vials (for 1 vial) Doxorubicin · HCl 0.8 g (10.0 mg)Polyvinylpyrrolidone 20.0 g (250.0 mg) Hydrochloric acid 0.1N pH = 3 (pH= 3) q.s. to Water for injections 0.4 l (5.0 ml) q.s. to

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.Polyvinylpyrrolidone was added and dissolved under stirring and nitrogenbubbling. The hydrochloric acid was then added dropwise to adjust the pHof the solution to 3. Further de-aerated water for injections was thenadded to bring the solution to its final volume (0.4 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 6:

TABLE 6 INITIAL VALUES Concentration: 1.973 mg/ml   pH = 2.71 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 2.028 102.8 1.944 98.5 1.791 90.8 1.477 74.9 21.885 95.5 1.582 80.2 0.972 49.3 3 1.840 93.2 1.402 71.0 0.632 32.0 41.913 97.0 1.853 93.9 1.273 64.5 8 1.972 99.9 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 5560 days t₉₀ at 8° C. =2670 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 7

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 8.00 g (100.0mg) N,N-Dimethylacetamide 0.12 l (1.5 ml) Hydrochloric acid 0.1N pH = 3(pH = 3) q.s. to Water for injections 0.40 l (5.0 ml) q.s. to

Doxorubicin.HCl (8.00 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-Dimethylacetamide was added under stirring and nitrogen bubbling.The hydrochloric acid was then added dropwise to adjust the pH of thesolution to 3. Further de-aerated water for injections was then added tobring the solution to its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 7:

TABLE 7 INITIAL VALUES Concentration: 19.32 mg/ml   pH = 2.96 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 20.1 103.5 19.14 99.1 17.34 89.8 15.57 80.6 219.20 99.4 15.77 81.6 12.94 67.0 3 18.06 93.5 14.85 76.9 11.61 60.1 420.03 103.7 17.81 92.2 13.78 71.3 8 19.99 103.5 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 1310 days t₉₀ at 8° C. =770 days

Similar stability data can be observed also for analogous solutionscontaining 4′-epi-doxorubicin, 4′-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 8

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.80 g (10.0 mg)Ethanol 0.12 l (1.5 ml) Hydrochloric acid 0.1N pH = 3 (pH = 3) q.s. toWater for injections 0.40 l (5.0 ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. Ethanol was addedunder stirring and nitrogen bubbling. Hydrochloric acid 0.1 N was thenadded dropwise to adjust the pH of the solution to 3. De-aerated waterfor injections was then added to bring the solution to its final volume(0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 12 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 8:

TABLE 8 INITIAL VALUES Concentration: 1.979 mg/ml   pH = 3.11 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 2.010 101.6 1.965 99.3 1.947 98.4 1.750 88.4 21.957 98.9 1.910 96.5 1.645 83.1 3 1.895 95.8 1.737 87.8 1.356 68.5 41.927 97.3 1.818 91.9 1.678 84.8 12 1.939 97.9 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 1270 days t₉₀ at 8° C. =780 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 9

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 8.000 g (100.00mg) N,N-Dimethylacetamide 0.060 l (0.75 ml) Propylene glycol 0.048 l(0.60 ml) Ethanol 0.012 l (0.15 ml) Hydrochloric acid 0.1N pH = 3 (pH =3) q.s. to Water for injections 0.400 l (5.00 ml) q.s. to

Duxorubicin.HCl (8.000 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-Dimethylacetamide, propylene glycol and ethanol were subsequentlyadded under stirring and nitrogen bubbling. The hydrochloric acid wasthen added dropwise to adjust the pH of the solution to 3. Furtherde-aerated water for injections was then added to bring the solution toits final volume (0.400 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials were tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 9:

TABLE 9 INITIAL VALUES Concentration: 20.07 mg/ml   pH = 2.99 Relative %Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 19.14 95.4 17.81 88.7 14.84 73.9 2 19.97 99.519.07 95.0 16.27 81.1 12.36 61.6 3 18.08 90.1 14.62 72.9 10.04 50.0 420.06 99.9 18.03 89.8 13.20 65.8 8 19.69 98.1 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 846 days t₉₀ at 8° C. =505 days

Similar stability data can be observed for analogous solutionscontaining 4′-epi-doxorubicin or 4′-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 10

for Composition 80 vials (for 1 vial) Doxorubicin · HCl 8.0 g (100.0 mg)Polyvinylpyrrolidone 20.0 g (250.0 mg) Hydrochloric acid 0.1N pH = 3 (pH= 3) q.s. to Water for injections 0.4 l (5.0 ml) q.s. to

Doxorubicin.HCl (8.0 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.Polyvinylpyrrolidone was added and dissolved under stirring and nitrogenbubbling. The hydrochloric acid was then added dropwise to adjust the pHof the solution to 3. Further de-aerated water for injections was thenadded to bring the solution to its final volume (0.4 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 10:

TABLE 10 INITIAL VALUES Concentration: 19.57 mg/ml   pH = 2.62 Relative% Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 19.54 99.9 19.11 97.6 16.88 86.2 12.48 63.8 218.43 94.2 14.13 72.2 6.00 30.7 3 18.02 92.1 11.57 59.1 2.61 13.3 419.58 100.1 17.36 88.7 9.23 47.2 8 19.34 98.8 t₉₀ (days) extrapolatedaccording to Arrhenius equation: t₉₀ at 4° C. = 2540 days t₉₀ at 8° C. =1290 days

Similar stability data can be observed for analogous solutionscontaining 4′-epi-doxorubicin or 4′-desoxy-doxorubicin, as hydrochloridesalts, at the same 20 mg/ml concentration.

EXAMPLE 11

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.80 g (10.0 mg)N,N-Dimethylacetamide 0.12 l (1.5 ml) Hydrochloric acid 0.1N pH = 3 (pH= 3) q.s. to Water for injections 0.40 l (5.0 ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling.N,N-Dimethylacetamide was added under stirring and nitrogen bubbling.Hydrochloric acid 0.1 N was then added dropwise to adjust the pH of thesolution to 3. De-aerated water for injections was finally added tobring the solution to its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 8 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 11:

TABLE 11 INITIAL VALUES Concentration: 1.826 mg/ml   pH = 3.14 Relative% Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.830 100.2 1.812 99.2 1.784 97.7 1.605 87.9 21.818 99.6 1.781 97.5 1.554 85.1 1.292 70.8 3 1.743 95.4 1.409 77.21.018 55.7 4 1.823 99.8 1.734 95.0 1.369 75.0 8 1.792 98.2 t₉₀ (days)extrapolated according to Arrhenius equation: t₉₀ at 4° C. = 5815 dayst₉₀ at 8° C. = 2920 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-epi-desoxy-4′-iodo-doxorubicin, daunorubicin or4-demethoxy-deunorubicin, as hydrochloride salts, at both 2 mg/ml and 5mg/ml concentration.

EXAMPLE 12

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.80 g (10.0 mg)Propylene glycol 0.12 l (1.5 ml) Hydrochloric acid 0.1N pH = 3 (pH = 3)q.s. to Water for injections 0.40 l (5.0 ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections de-aerated by nitrogen bubbling. Propylene glycolwas added under stirring and nitrogen bubbling. Hydrochloric acid 0.1 Nwas then added dropwise to adjust the pH of the solution to 3.De-aerated water for injections was finally added to bring the solutionto its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 4 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 12:

TABLE 12 INITIAL VALUES Concentration: 1.982 mg/ml   pH = 3.11 Relative% Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.972 99.5 1.934 97.6 1.889 95.3 1.705 86.0 21.952 98.5 1.795 90.6 1.483 74.8 3 1.935 97.6 1.699 85.7 1.153 58.2 42.056 103.7 1.788 90.2 1.460 73.7 t₉₀ (days) extrapolated according toArrhenius equation: t₉₀ at 4° C. = 1794 days t₉₀ at 8° C. = 1025 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml 5 mg/ml concentration.

EXAMPLE 13

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.80 g (10.0 mg)Polyethylene glycol 400 0.12 l (1.5 ml) Hydrochloric acid 0.1N pH = 3(pH = 3) q.s. to Water for injections 0.40 l (5.0 ml) q.s. to

Doxorubicin.HCl (0.80 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. Polyethyleneglycol 400 was added under stirring and nitrogen bubbling. Hydrochloricacid 0.1 N was then added dropwise to adjust the pH of the solution to3. De-aerated water for injections was finally added to bring thesolution to its final volume (0.40 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 55° C., 45° C. and 35° C. (acceleratedstability controls) and at 4° C. for up to 3 weeks (55° C.), 4 weeks(45° C. and 35° C.) and 4 weeks (4° C.).

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 13:

TABLE 13 INITIAL VALUES Concentration: 1.907 mg/ml   pH = 3.07 Relative% Assay: 100.0 TEMPERATURE 4° C. 35° C. 45° C. 55° C. TIME Conc. Rel. %Conc. Rel. % Conc. Rel. % Conc. Rel. % (weeks) mg/ml Assay mg/ml Assaymg/ml Assay mg/ml Assay 1 1.871 98.1 1.797 94.2 1.668 87.5 1.484 77.8 21.710 89.7 1.608 84.3 1.237 64.9 3 1.739 91.2 1.551 81.3 1.007 52.8 41.873 98.2 1.693 88.8 1.453 76.2 t₉₀ (days) extrapolated according toArrhenius equation: t₉₀ at 4° C. = 1130 days t₉₀ at 8° C. = 680 days

Similar stability data can be observed also for analogous solutionscontaining either doxorubicin hydrochloride at 5 mg/ml concentration, or4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin or 4-demethoxy-daunorubicin,as hydrochloride salts, at both 2 mg/ml and 5 mg/ml concentration.

EXAMPLE 14

Composition for 80 vials (for 1 vial) Doxorubicin · HCl 0.8 g (10 mg)Hydrochloric acid 0.1N pH = 3 (pH = 3) q.s. to Water for injections 0.4l (5 ml) q.s. to

Doxorubicin.HCl (0.8 g) was dissolved in 90 percent of the amount ofwater for injections, de-aerated by nitrogen bubbling. The hydrochloricacid was then added dropwise to adjust the pH of the solution to 3.Further de-aerated water for injections was then added to bring thesolution to its final volume (0.4 l).

The solution was filtered through a 0.22 μm microporous membrane undernitrogen pressure. Volumes of 5 ml of the solution were distributed intotype I-colourless glass vials having 5/7 ml capacity. The vials werethen closed with chlorobutyl teflon-faced rubber stoppers and sealedwith aluminium caps.

The stability of the solutions in the vials was tested. The vials werestored at temperatures of 4° C. and 8° C. for up to 6 months.

The stability data obtained, using high performance liquidchromatography (HPLC) for the determination of potency, are reported inthe following Table 14:

TABLE 14 INITIAL VALUES Concentration: 2.039 mg/ml pH = 3.06 Relative %Assay: 100.0 TEMPERATURE 4° C. 8° C. TIME Conc. Rel. % Conc. Rel. %(weeks) mg/ml Assay mg/ml Assay 1 1.983 97.3 1.959 96.1 3 1.984 97.31.983 97.3 6 2.012 98.7 2.002 98.2

At the same conditions, similar stability data can be generally observedalso for the other solutions mentioned in the preceding examples.

We claim:
 1. A sterile, pyrogen-free, anthracycline glycoside solutionwhich consists essentially of a physiologically acceptable salt of ananthracycline glycoside dissolved in a physiologically acceptablesolvent therefor, which has not been reconstituted from a lyophilizateand which has a pH of from 2.5 to 6.5.
 2. A solution according to claim1 in a sealed container.
 3. A solution according to claim 1 or 2,wherein the anthracycline glycoside is selected from the groupconsisting of doxorubicin, 4′-epi-doxorubicin, 4′-desoxy-doxorubicin,4′-desoxy-4′-iodo-doxorubicin, daunorubicin and4-demethoxy-daunorubicin.
 4. A solution according to claim 3, whereinthe anthracycline glycoside is doxorubicin.
 5. A solution according toclaim 1, wherein the physiologically acceptable salt of theanthracycline glycoside is the salt with a physiologically acceptableacid selected from the group consisting of hydrochloric, hydrobromic,sulfuric, phosphoric, nitric, acetic, succinic, tartaric, ascorbic,citric, glutamic, benzoic, methanesulfonic and ethanesulfonic acid.
 6. Asolution according to claim 5, wherein the physiologically acceptablesalt of the anthracycline glycoside is the salt with hydrochloric acid.7. A solution according to claim 1, having a pH of from 2.5 to 5.5.
 8. Asolution according to claim 7, having pH 3 or pH
 5. 9. A solutionaccording to claim 1, wherein the physiologically acceptable solvent forthe anthracycline glycoside is selected from the group consisting ofwater, ethanol, polyethyleneglycol, dimethylacetamide and mixturesthereof.
 10. A solution according to claim 9, wherein thephysiologically acceptable solvent is water.
 11. A solution according toclaim 1, wherein the concentration of the anthracycline glycoside isfrom 0.1 mg/ml to 100 mg/ml.
 12. A solution according to claim 11,wherein the concentration of the anthracycline glycoside is from 0.1mg/ml to 50 mg/ml.
 13. A solution according to claim 12, wherein theconcentration of the anthracycline glycoside is from 1 mg/ml to 20mg/ml.
 14. A sterile, pyrogen-free, doxorubicin solution which consistsessentially of a physiologically acceptable salt of doxorubicindissolved in a physiologically acceptable solvent therefor, which hasnot been reconstituted from a lyophilizate and which has a pH of from2.5 to 6.5.
 15. A solution according to claim 14 in a sealed container.16. A solution according to claim 14, wherein the physiologicallyacceptable salt of doxorubicin is the salt with a physiologicallyacceptable acid selected from the group consisting of hydrochloric,hydrobromic, sulfuric, phosphoric, nitric, acetic, succinic, tartaric,ascorbic, citric, glutamic, benzoic, methanesulfonic and ethanesulfonicacid.
 17. A solution according to claim 16, wherein the physiologicallyacceptable salt of doxorubicin is the salt with hydrochloric acid.
 18. Asolution according to claim 14, having a pH of from 2.5 to 5.5.
 19. Asolution according to claim 18, having pH 3 or pH
 5. 20. A solutionaccording to claim 14, wherein the physiologically acceptable solventfor doxorubicin is selected from the group consisting of water, ethanol,polyethyleneglycol, dimethylacetamide and mixtures thereof.
 21. Asolution according to claim 20, wherein the physiologically acceptablesolvent is water.
 22. A solution according to claim 14, wherein theconcentration for doxorubicin is from 0.1 mg/ml to 100 mg/ml.
 23. Asolution according to claim 22, wherein the concentration of doxorubicinis from 2 mg/ml to 50 mg/ml.
 24. A solution according to claim 23,wherein the concentration of doxorubicin is 2 mg/ml to 20 mg/ml.
 25. Asolution according to claim 24, wherein the concentration of doxorubicinis 2 mg/ml or 5 mg/ml.
 26. A process for producing the solution of claim1, which process comprises dissolving a physiologically acceptable saltof the anthracycline glycoside, which salt is not in the form of alyophilizate, in a physiologically acceptable solvent therefor;optionally adding a physiologically acceptable acid or buffer to adjustthe pH within the said range as desired; and passing the resultingsolution through a sterilising filter.
 27. A process according to claim26, wherein an additional component selected from the group consistingof co-solubilizing agents, tonicity adjustment agents and preservativesis added to the solution prior to passing the solution through asterilizing filter.
 28. A process according to claim 27, wherein theco-solubilizing agent is polyvinylpyrrolidone.
 29. A method ofinhibiting the growth of a tumour selected from the group consisting ofsarcomas, carcinomas, lymphomas, neuroblastoma, melanoma, myeloma,leukemias and Wilms tumour, which method comprises administering to ahost suffering from said tumour the solution of claim 1, containing theactive drug substance in an amount sufficient to inhibit the growth ofsaid tumour.
 30. A method according to claim 29, wherein the tumour isselected from the group consisting of Moloney Sarcoma Virus, Sarcoma 180Ascites, Solid Sarcoma 180, gross transplantable leukemia, L 1210leukemia and lymphocitic P 388 leukemia.